To establish a method of directional differentiation and efficient production of neurons from embryonic stem cells (ES cells) in vitro, based on the 4-/4+ protocol described by Bain, a new method was established to induce ES cells differentiating into neurons by means of three-step differentiation using all-trans retinoic acid (ATRA) combined with astrocyte-conditioned medium (ACM) in Vitro. The totipotency of ES cells was identified by observation of cells' morphology and formations of teratoma in immunocompromised mice. The cells' differentiation was evaluated continuously by the detection of the specific cellular markers of neural stem cells, neurons and astrocytes, including nestin, NSE and GFAP using immunohistochemistry assay. The NSE positive cells' ratio of the differentiated cells was determined by flow cytometry. It was found that the transparent circular clusters surrounding embryoid bodies induced with combining induction protocol formed just after 24 h and gradually enlarged later. This phenomenon could not be observed in EBs induced only by ATRA. The NSE positive cells' ratio in the cells induced with ATRA and ACM was higher than that of the cells induced by ATRA at different time points of differentiation, and finally reached up to 73.5% among the total differentiated population. It was concluded that ES cells could be induced into neurons with high purity and yield by means of inducing method combining with ATRA and ACM.
Astrocytes/*cytology ; Cell Differentiation ; Cells, Cultured ; Embryo, Mammalian ; Neurons/*cytology ; Stem Cells/*cytology ; Tretinoin/pharmacology

Background Interleukin-17 (IL-17)is a potent pro-inflammatory cytokine and plays a pathogenic role in autoimmune disease.It was confirmed that IL-17 is implicated in allograft rejection of many transplanted organs.Recent studies have foensed on the effect of IL-17 antagonists on allograft rejection.Objective This study aimed to investigate the inhibitory effect of anti-mouse IL-17 monoclonal antibody (mAb) on corneal allograft rejection.Methods Twenty-five 8 to 10-week-old C57BL/6 mice and 50 BALB/c mice were collected.Donor cornea grafts with 2 mm diameter from 25 C57BL/6 mice was transplanted to 50 eye of BALB/c mice to establish a model of corneal transplantation.The recipients were randomized into 2 groups,and neutralizing mouse IL-17antibody or isotype control antibody was intraperitoneally injected immediately after transplantation for experimental treatment,respectively.Allografts were scored clinically at appropriate time points after treatment based on Plskova criteria,and ≥5 was confirmed as rejection.Infiltrating cells in corneal graft were detected qualitatively and quantitatively by immunohistochemistry and reverse transcription-PCR separately.The cytokine levels of T helper type 1 (Th1),Th2,and Th17 in recipients' spleen wer(c) analyzcd by ELISA.The use of the animals followed the Statement of ARVO.Results Compared with the isotype control antibody group,the survival of grafts was improved in the IL-17mAb group(P＜0.05).The levels of neutrophile granulocyte mRNA,CD4+ and CD8+ T lymphotes mRNA were 2.22±0.10,1.64±0.04 and 1.32±0.10 in the IL-17 mAb group,showing a significant decline in comparison with those of the isotype control antibody group(3.61 ±0.08,2.69±0.06 and 2.17±0.04) (P=0.000,0.000,0.000).Interferon-γ(IFN-γ),IL-12 p40 and IL-17 concentrations in recipients ' splenocytes were (529.80 ± 13.83) ng/L,(539.58 ±10.74) ng/L and(173.70±8.11)ng/L in the IL-17 mAb group,and thosc in the isotype control antibody group were (741.48± 10.51) ng/L,(1156.90 ± 69.93) ng/L and (366.13± 7.93) ng/L,with significant differences between them (P=0.000,0.001,0.000).Conclusions Neutralization IL-17 bioactivity inhibits mouse corneal allograft rejection to a certain extent.

Objective To determine whether homocysteine(Hcy)and monosodium glutamate (MSG)could lead to animal model of hypertensive disorder complicating pregnancy and its mechanism. Methods Female adult Wistar rats were randomly divided into 4 groups:pregnant control group(PN), pregnant Hcy group(PH),pregnant glutamic acid group(PG)and pregnant Hcy and glutamie acid group (PHG).The rats of each group were injected with Hey 200 mg/kg or physiological saline every day intraperitoneally and with MSG or 0.9% saline every other day via Hcy injection from the 10th day to the 20th day of pregnancy.The blood pressure,urine protein,function of liver and kidney,weight of placenta, length and weight of fetus were all measured.The histological change of the pallium and the change of behavior of pregnant rats were also observed.Results(1)The blood pressure in PH[(107?8)mm Hg, 1 mm Hg=0.133 kPa],and PHG group [(109?10)mm Hg] after the treatment increased significantly compared with those in other groups from the 12 th day after pregnancy(P

Saccharmyces cerevisiae 220 has certain resistance to Pb2+ was found through the test of it's cultured on media containing lead cation. Sacchannyces cerevisiae 220 cultured was put into the media cotaining lead cation and it's biosorp-tion to lead cation was studied. The results show this strain has maximum biosorption ratio (96.6%) to 6mg/L lead cation, biosorption-equilibrilium period is 25-30min, biosorption kinitics equation is q-26.318C/ (1 + 0.437C) . Acetic acid is benefit to de-biosorption, glucose and KH2PO4 can improve biosorption ratio to lead cation.

A.niger M1, the initial strain, was treated by UV and a mutant with 30% higher xylanase activity was obtained. Zymogram for detecting xylanase showed there are three different xylanases in the mutant mature culture, while two xylanases in initial strain. After orthogonal experiment, the optimum fermentation conditions of the mutant were obtained as follows: concentration of the major carbon resource 4 %, ratio between bran and corncob 5:5, concentration of glucose 0.1%, concentration of ammonium oxalate as supplemental nitrogen resource 2.0%, the initial pH of liquid medium 5.0, 100mL/250mL flask.

Objective To investigate the prevalence of clonorchiasis among residents of Huachuan Country,Heilongjiang Province and to provide a basis for development of control strategies.Methods From 2011 to 2012,cluster random sampling was performed to survey the incidence of clonorchiasis in Huachuan Country.Fecal specimens were collected and examined the clonorchis sinensis eggs by Kato-Katz method.A questionnaire survey was conducted to collect related information such as age,gender,occupation and eating habits.The infection characteristic was analyzed.Results Totally 884 patients with clonorchiasis were found among 2248 residents,and the infection rate was 39.32％.The infection rate in male[47.15％(611/1296)] was significantly higher than females [28.68％(273/952),x2 =34.55,P ＜ 0.01].The infection rate increased with age,which was higher in the 20-69 years old people,with the highest infection rate in the 50-59 years old groups[45.34％ (219/483)].Of the occupational distribution,farmers had the highest infection rate [47.24％ (420/889)],followed by cadres and staffs[38.38％(190/495)].Of residents with fresh fish eating history,the prevalence of clonorchiasis was 53.38％(150/281).Conclusions The prevalence of clonorchiasis is still high in Huachuan County.To reduce the prevalence of clonorchiasis,comprehensive prevention measures,health education and group chemotherapy should be carried out.

Aged ; Fatal Outcome ; Heart Rupture ; etiology ; Humans ; Male ; Myocardial Infarction ; therapy

OBJECTIVETo investigate clinicopathological and genetic characteristics of primary ocular adnexal lymphoproliferative lesions.

METHODSClinical, morphological and immunohistochemical features of 37 archival cases of primary ocular adnexal lymphoproliferative lesions were studied including 5 cases of reactive lymphoid hyperplasia and 32 lymphomas retrospectively. Classification of the lymphomas were made according to the WHO classification of tumors of haematopoietic and lymphoid tissues. All cases were studied by interphase fluorescence in situ hybridization (FISH) using dual color break apart probes of IgH, MALT1, bcl-6, c-Myc, bcl-2, CCND1, bcl-10, and FOXP1 for detection of chromosomal aberrations involving IgH, MALT1, bcl-6, c-Myc, bcl-2, cyclinD1, bcl-10 and FOXP1 genes, respectively. FISH with IgH / bcl-2 dual color dual fusion probe was used for detection of t(14;18)(q32;q21)/IgH-bcl-2. CEP18 spectrum orange probe was used for detection of aneuploidy of the chromosome 18.

RESULTSAmong 32 cases of lymphomas, 28 cases (87.5%) were extranodal marginal zone B-cell lymphomas of mucosa associated lymphoid tissue (MALT lymphoma), 2 cases were follicular lymphoma (FL) and 2 cases diffuse large B cell lymphoma (DLBCL). Among the 28 cases of MALT lymphoma, chromosomal aberrations were found in 60.7% (17/28) by interphase FISH analysis. One case showed positive IgH break-apart signal with unknown partner. 16 cases showed three copies of different genes, of which, three copies of MALT1, bcl-6, and c-Myc were identified in 7 cases (25%), 12 cases (43%), and 2 cases (8%) of MALT lymphomas, respectively. In addition, 5 cases showed two genes including three copies of bcl-6 and MALT1 in 4 cases, and three copies of bcl-6 together with c-Myc in one case. Furthermore, all cases with three copies of MALT1 had trisomy 18. t(14;18)(q32;q21) was detected in both follicular lymphomas. Of the 2 DLBCL cases, one showed three copies of bcl-6 together with trisomy 18 and the other one showed three copies of bcl-6 together with IgH and c-Myc rearrangements. Chromosomal aberration was not found in all 5 cases of reactive lymphoid hyperplasia.

CONCLUSIONSThe most common entity of primary ocular adnexal lymphomas is MALT lymphoma and FISH is helpful for their differential diagnosis and classification. Trisomy 18 and three copies of bcl-6 are common chromosomal aberrations in primary ocular adnexal MALT lymphomas.

Aneuploidy ; B-Lymphocytes ; pathology ; Caspases ; genetics ; Chromosome Aberrations ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 18 ; Eye ; pathology ; Eye Neoplasms ; genetics ; pathology ; Female ; Genes, bcl-2 ; genetics ; Humans ; Immunoglobulin Heavy Chains ; genetics ; In Situ Hybridization, Fluorescence ; Interphase ; Lymphoma, B-Cell ; genetics ; Lymphoma, B-Cell, Marginal Zone ; genetics ; Lymphoma, Large B-Cell, Diffuse ; genetics ; pathology ; Male ; Mutation ; Translocation, Genetic ; Trisomy

6.24 mmol/L) and sixty healthy persons in the health center of our hospital were investigated as hyperhpidemia group (Hyperlipidemias) and control group (Controls) respectively.Hyperlipidemias were given simvastatin 20 mg?d~(-1) for twelve weeks (Statins).Blood samples of ulnar vein were extracted from Statins at the end of twelve weeks as well as Controls and Hyperhpidemias at the beginning of the experiment. Blood serum,plasma and mononuclearcell were extracted and stored at a refrigerator of-80℃.The level of plasma angiotensinⅡwas detected by the method of radioimmunity.While the expression of RANTES mRNA and protein on mononuclearcell were assessed by real time reverse transcription polymerse chain reaction and Western blot respectively.Results①The plasma angiotensinⅡof Hyperlipidemias was higher than that of Controls [(92.13?22.03) vs (50.85?12.12),P

Background and Objective Keshan disease is clinically characterized as a dilated eardiomyopathy. We analyze the prevalence trend during last decade of hypertension and Keshan-disease in Yangzhuang village which was a Keshan-disease epidemic area.Method The survey including medical history,blood pressure and ECG were carried out every two years during the follow up 13 years.Results During follow up period,the total detection rate(hypertension:13.4 % vs Keshan-disease:10.7 %,?~2=8.555,P=0.002)and the accumulative rate of hypertension were higher than those of Keshan-disease,which was on the contrary to that before 1993,when increasing rate of Keshan-disease was higher than hypertension.Furthermore,the accumulative increasing rate of hypertension was 240.0%,which was higher than the national average level during corresponding period with no significant differences between female and male.Conclusion The detection rate of hypertension in Keshan disease epidemic area was higher than the average rate nation-wide.Whether the hypertension prevalence was re- lated to Keshan-disease needs further investigation.